Genomic semi-automated cycle sequencing as a sensitive screening technique for p53 mutations in frozen tumor samples.
Single-strand conformational polymorphism (SSCP) is the most widely used method for p53 gene mutation screening. Nucleotide sequence analysis is considered more sensitive for detection of mutations. We established a genomic semi-automated cycle sequencing protocol suitable for p53 gene mutation screening. The technique was applied to 44 SSCP-negative frozen ovarian cancer samples: Eleven mutations (11/44, 25%) were found, 6 point missense mutations, 3 silent point mutations, 1 nonsense mutation and 1 single-base deletion. Heterozygous mutations were readily detectable. Genomic semi-automated cycle sequencing is a sensitive, time-effective screening method requiring only small amounts of tumor tissue.
Wang-Gohrke
S
S
Department of Obstetrics and Gynecology, Molecular Biology Laboratory, University of Ulm, Germany.
Hees
S
S
Pochon
A
A
Wen
W H
WH
Reles
A
A
Press
M F
MF
Kreienberg
R
R
Runnebaum
I B
IB
D019943:Amino Acid Substitution; D001331:Automation; D018389:Codon, Nonsense; D005260:Female; D016368:Frameshift Mutation; D016158:Genes, p53; D006801:Humans; D009154:Mutation; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000473:pathology*; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D018807:Polymorphism, Single-Stranded Conformational; D012680:Sensitivity and Specificity; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry / Q000235:genetics
D018389:Codon, Nonsense; D016159:Tumor Suppressor Protein p53